• Title/Summary/Keyword: pathogenic Escherichia coli

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Genotyping of avian pathogenic Escherichia coli by DNA fragment analysis for the differences in simple sequence repeats

  • Han, Mi Na;Byeon, Hyeon Seop;Han, Seong Tae;Jang, Rae Hoon;Kim, Chang Seop;Choi, Seok Hwa
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.257-262
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    • 2018
  • Avian pathogenic E. coli (APEC) causes severe economic losses in the poultry farms, due to systemic infections leading to lethal colisepticemia. It causes a variety of diseases from air sac infection to systemic spread leading to septicemia. Secondary infection contains opportunistic infections due to immunosuppression disease. Collibacillosis causes the great problems in the poultry industry in Korea. Thus, it is necessary to identify and classify the characteristics of E. coli isolate of chicken origin to confirm the diversity of symptoms and whether they are transmitted among the farms. Fragment analysis is identify the difference in the number of Variable-Number Tandem-Repeats (VNTRs) for genotyping. VNTRs have repeating structure (Microsatellite, Short tandem repeats; STR, Simple sequence repeats; SSR) in the chromosome. This region can be used as a genetic marker because of its high mutation rate. And various lengths of the amplified DNA fragment cause the difference in the number of repetition of the DNA specific site. The number of repetition sequences indicates the separated size of fragments, so the each fragments can be distinguished by specific samples. The results of the sample show that there is no difference in six microsatellite loci (yjiD, aidB, molR_1, ftsZ, b1668, yibA). There are differences among the farms in relation of the number of repetitions of other six microsatellite loci (ycgW, yaiN, yiaB, mhpR, b0829, caiF). Four (ycgW, yiaB, b0829, caiF) of these six microsatellite loci show statistically significant differences (P<0.05). It means that the analysis using four microsatellite loci including ycgW, yiaB, b0829, and caiF can confirm among the farms. Five E. coli samples in one farm have same SSR repetition at all markers. But, there are significant differences from other farms at Four (ycgW, yiaB, b0829, caiF) microsatellite loci. These results emphasize again that the four microsatellite loci makes a difference in the amplified DNA fragments, enabling it to be used for E. coli genotyping.

Efficient Treatment Methods for Reducing Escherichia coli Populations in Commercially-Available Red Pepper Powder in Korea (국내 유통 고춧가루의 병원성 대장균 오염 및 대장균 저감화 방법)

  • Song, Young-Jin;Park, Se-Won;Chun, Se-Chul;Choi, Mi-Jung;Chung, Koo-Chun;Lee, Si-Kyung
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.6
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    • pp.875-880
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    • 2012
  • This study was conducted to investigate the level of contamination of pathogenic Escherichia (E.) coli in 50 types of red pepper powders collected domestically. Pathogenic E. coli was confirmed using real-time PCR to confirm the 4 types of EAEC, EPEC, EHEC and ETEC. One sample out of 50 was contaminated with pathogenic E. coli. The type of pathogenic E. coli detected in the sample was EAEC. This study was also conducted to determine the effect of alcohol treatment on the reduction of E. coli populations in red pepper powder. The amount of E. coli in the control was $1.2{\times}10^6$ cfu/mL. The amount of E. coli in 10 minutes immersion treatment with 10% alcohol was $1.1{\times}10^6$ cfu/mL. In samples treated with over 20% alcohol, E. coli was not detected. This showed that 10 minutes of immersion in over 20% alcohol might be effective to reduce E. coli. This study was also conducted to determine the effect of UV irradiation on E. coli reduction. The number of E. coli in the control group was $5.0{\times}10^5$ cfu/mL. However, the number of E. coli in 45 min of the UV irradiated sample decreased to $1.0{\times}10^3$ cfu/mL, by $10^2$ cfu/mL. In contrast, E. coli was not detected in an over 60 min UV irradiated sample in $10^{-2}dilution$. This study showed that over 20% alcohol treatment and UV irradiation for 60 min was effective to control E. coli in red pepper powder.

Random amplified polymorphic DNA analysis of bacterial pathogens using universal rice primers

  • Monoldorova, Sezim;Kim, Jinsol;Kim, Joon Hee;Jeon, Bo-Young
    • Korean Journal of Veterinary Service
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    • v.40 no.1
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    • pp.1-6
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    • 2017
  • Molecular typing of pathogenic microorganisms is important for epidemiological investigation of infectious disease outbreaks. In this study, we applied Universal Rice Primers (URP) that were originated from repetitive sequences in rice chromosomal DNA to random amplified polymorphic DNA (RAPD) analysis of pathogenic bacteria such as Escherichia coli, Listeria monocytogenes, and Salmonella sp. Of the twelve URP primers examined to date, seven primers (URP-2, -3, -4, -5, -6, -8, and -9) generated reproducible and polymorphic PCR products ranging from 1 to 13 bands. One of them, URP-6 was very effective in differentiating seven E. coli serotypes, seven L. monocytogenes clinical isolates, and eight Salmonella subspecies (ssp.) serovars. The results thus indicate that RAPD analysis using URP primers might be useful in typing bacterial pathogens including E. coli, L. monocytogenes, and Salmonella strains.

Antibiotic Resistance and Assessment of the Food-borne Pathogenic Microorganisms in Ready to Eat Meals (식사대용 식품(Ready-to-eat meals) 중 병원성 세균의 분포와 항생제 감수성 양상)

  • Hong, Eun-Kyung;Kim, Yun-A;Lee, Do-Kyung;Kang, Byung-Yong;Ha, Nam-Joo
    • Korean Journal of Microbiology
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    • v.42 no.4
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    • pp.265-270
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    • 2006
  • This study was performed in order to measure the level of food-borne pathogenic bacteria and antibiotic resistance pattern of found ready to eat meals such as Him-bap, Cho-bap, Hamburger, Sandwich and packed lunch boxes. A total of 497 samples were collected from supermarket and department of Seoul, Kyung-ki, Inctleon, Kang-won, Chung-Cheong from November, 2005 to March, 2006. The contaminated microorganisms were in most cases tract relative strain like E. coli and S. aureus. Result have shown E. coli was detected 4 strains and S. aureus was detected 22 strains. 26 strains were also tested the antibiotic resistance pattern. 26 strains were shown to be relatively susceptible to synercid, vancomycin, teicoplanin, ciprofloxacin, gentamycin, lincomycn, cefotaxime, meropenem, cephalosporin, amoxicillin-clavulanic acid by the MIC dilution method, but E. coli 1 strain was resistant to amoxicillin-clavulanic acid.

Establishment of a linear regression equation for quantification of beta-hemolytic Escherichia coli in different media and survival of hemolytic Escherichia coli after blending with three different media

  • Kim, Jae Cheol;Pluske, John R.;Yoo, Jaehong;Heo, Jung Min
    • Korean Journal of Agricultural Science
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    • v.41 no.2
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    • pp.135-139
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    • 2014
  • Pathogenic E. coli associated post-weaning diarrhea (PWD) and edema disease are common diseases in commercially-housed weanling pigs. An enterotoxigenic E. coli (ETEC) oral challenge model has been used to mimic the physiological responses observed in commercial conditions. However, an oral challenge procedure has two major limitations: (1) the ETEC cell density is unknown at the point of oral inoculation, and (2) blending ETEC with traditional TSB (trypticase soy broth) is not palatable and hence decreases acceptability by piglets. Therefore, the purposes of this study were to (1) establish a regression equation that can be used for estimation of ETEC concentration in dilution media using the spectrophotometric measurement of cell density; and (2) examine survival of ETEC after blending either with TSB, sweetener or dextrose. A strain of ETEC (serogroup beta-hemolytic E. coli O149; K91; F4; toxins LT, STa, STb) was grown in TSB for 3.5 hours, centrifuged, the supernatant was discarded, and the ETEC pellet was then blended either with TSB (100 mL), sweetener (60 mL TSB + 40 mL fruit flavored concentrate), or dextrose (50 mL TSB + 50 mL dextrose; 0.5g/mL dextrose). Cell density was measured using the colorimetric method and also plated on a 5% sheep blood agar for counting of ETEC colony forming units at 0, 5, 35, 65 and 125 min after blending. The optical density at 600 nm explained 83% of ETEC colony forming units, indicating that the established linear equation (y= 6E+08x - 4E+07, P<0.004) can be used for robust quantification of ETEC cell density in TSB, sweetener and dextrose media. When ETEC was blended with sweetener and dextrose, survival of ETEC was decreased by 45% and 72% within 5 min post-blending. Therefore, further research is required to find out the suitable medium that has potential to improve palatability without compromising survival of ETEC.

Studies on the pathogenic Escherichia coli isolated from piglets with diarrhea in Chonnam (전남 지방의 설사 자돈에서 분리된 병원성 대장균에 관한 연구)

  • 김자숙;박영춘;정인호;오은희;박석준;고홍범
    • Korean Journal of Veterinary Service
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    • v.19 no.2
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    • pp.139-153
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    • 1996
  • Porcine E coli infection is a disease caused by Enterotoxin produced by Enterotoxigenic Escherichia coli(ETEC). Enteric colibacillosis has become an economically important disease in pigs as a result of increasing intensification of farrowing management. The present study undertaken to obtain the antibiotic sensitivity and distribution of serogroups and pili producibility test of ETEC from E. coli isolates in Chonnam. The results obtained were as follows. 1. A total of 71 isolates identified as E, coli employing IMViC system from rectal specimens of 54 piglets with diarrhea. 2. In antibiotic sensitivity test, isolates showed high sensitivity to AN, CM, Fox, GM, but resistance to EM, NA TC. 3. The distribution of 25 Isolates of serogroups were 0141:K85(11.3%), 08:K87(8.5%), 064:K (5.6%), 0138:K8l (4.2%), 0139 :K82(2.8%), 0157:K88ac(1.4%) and 0149:K9l (1.4%). 4. MRHA of guinea pig erythrocytes was detected in 8 out of 25OK serotypes and 9 out of 46 unidentified serotypes. MRHA titers of serotypes showed from 64 to 128 in 0141: K85, 2 in 0138:K8l and no titers in 0139:K82. 5. The production of heat labile enterotoxin of ETEC was detect 39 out of 52 isolates showed $\beta$-hemolysin, 7 out of 52 isolates showed ${\gamma}$-hemolysin and 6 out of 52 isolates showed ${\gamma}$-hemolysin by $GM_1$ganglioside ELISA. The distribution of LT toxin were in 12 isolate showed $\beta$-hemolysin, 2 isolates showed ${\alpha}$-hemolysin and 3 isolates showed ${\gamma}$-hemolysin in 25 OK serotypes.

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Two Enteropathogenic Escherichia coli Strains Representing Novel Serotypes and Investigation of Their Roles in Adhesion

  • Wang, Jing;Jiao, HongBo;Zhang, XinFeng;Zhang, YuanQing;Sun, Na;Yang, Ying;Wei, Yi;Hu, Bin;Guo, Xi
    • Journal of Microbiology and Biotechnology
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    • v.31 no.9
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    • pp.1191-1199
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    • 2021
  • Enteropathogenic Escherichia coli (EPEC), which belongs to the attaching and effacing diarrheagenic E. coli strains, is a major causative agent of life-threatening diarrhea in infants in developing countries. Most EPEC isolates correspond to certain O serotypes; however, many strains are non-typeable. Two EPEC strains, EPEC001 and EPEC080, which could not be serotyped during routine detection, were isolated. In this study, we conducted an in-depth characterization of their putative O-antigen gene clusters (O-AGCs) and also performed constructed mutagenesis of the O-AGCs for functional analysis of O-antigen (OAg) synthesis. Sequence analysis revealed that the occurrence of O-AGCs in EPEC001 and E. coli O132 may be mediated by recombination between them, and EPEC080 and E. coli O2/O50 might acquire each O-AGC from uncommon ancestors. We also indicated that OAg-knockout bacteria were highly adhesive in vitro, except for the EPEC001 wzy derivative, whose adherent capability was less than that of its wild-type strain, providing direct evidence that OAg plays a key role in EPEC pathogenesis. Together, we identified two EPEC O serotypes in silico and experimentally, and we also studied the adherent capabilities of their OAgs, which highlighted the fundamental and pathogenic role of OAg in EPEC.

Detection of CTX-M and Clonal Analyses using MLST in Cefotaxime Resistant $Escherichia$ $coli$ Isolated from the Han-River, Korea

  • Son, Gun-Woo;Kim, Mal-Nam
    • Korean Journal of Environmental Biology
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    • v.29 no.4
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    • pp.286-295
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    • 2011
  • Bacteria resistant to various antibiotics have recently become an issue of the utmost importance. Resistant strains are not uncommon, even in municipal drinking water sources. The health threat posed by resistant, pathogenic bacteria has serious ramifications for both public health and agriculture. In this study, we isolated antibiotic resistant bacteria from water samples from the Han River, Korea, which is contaminated by the wastewater from many industrial complexes, hospitals, agricultural and animal husbandry estates, and from wastewater treatment facilities. We determined the degrees of resistance to various antibiotics exhibited by the isolated strains. The similarities between the isolated $E.$ $coli$ strains were examined, using the pulsed field gel electrophoresis and multilocus sequence typing, in order to trace their origins and to explore the syntechnic adaptations and pathogenicity of the various strains and relate these to their genetic sequence. A total of 25 $E.$ $coli$ strains were isolated from six stations along the Han River. All the 25 strains exhibited resistance to ampicillin. We also investigated resistance to amoxicillin, clavulanic acid, cefazolin, cofoxitin, cefotaxime, cefpodoxime, ceftriaxone, cefepime, nalidixic acid, aztreonam, ciprofloxacin, streptomycin, gentamicin, chloramphenicol and imipenem. Based on the ESBL detection, 14 strains belonged to the ESBL producing strains. The number of the clonal complex producing strains was 5 among the 14 isolated strains. The 5 strains were included in the 168, 23, 38, 469, 156 clonal complex, respectively. The rest 9 strains were not included in the clonal complex, but showed independent STs.

Detection of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes in Kimchi by Multiplex Polymerase Chain Reaction (mPCR)

  • Park, Yeon-Sun;Lee, Sang-Rok;Kim, Young-Gon
    • Journal of Microbiology
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    • v.44 no.1
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    • pp.92-97
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    • 2006
  • We developed an mPCR assay for the simultaneous detection, in one tube, of Escherichia coli O157:H7, Salmonella spp., Staphylococcus aureus and Listeria monocytogenes using species-specific primers. The mPCR employed the E. coli O157:H7 specific primer Stx2A, Salmonella spp. specific primer Its, S. aureus specific primer Cap8A-B and L. monocytogenes specific primer Hly. Amplification with these primers produced products of 553, 312, 405 and 210 bp, respectively. All PCR products were easily detected by agarose gel electrophoresis, and the sequences of the specific amplicons assessed. Potential pathogenic bacteria, in laboratory-prepared and four commercially available kimchi products, were using this mPCR assay, and the amplicons cloned and sequenced. The results correlated exactly with sequences derived for amplicons obtained during preliminry tests with known organisms. The sensitivity of the assay was determined for the purified pathogen DNAs from four strains. The mPCR detected pathogen DNA at concentrations ranging from approximately 0.45 to $0.05\;pM/{\mu}l$. Thus, this mPCR assay may allow for the rapid, reliable and cost-effective identification of four potentially pathogens present in the mixed bacterial communities of commercially available kimchi.

The inhibitory effect of natural bioactives on the growth of pathogenic bacteria

  • Kim, Ji-Sun;Kim, Yang-Ha
    • Nutrition Research and Practice
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    • v.1 no.4
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    • pp.273-278
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    • 2007
  • The objective of this study was to evaluate the inhibitory activity of natural products, against growth of Escherichia coli (ATCC 25922) and Salmonella typhimurium (KCCM 11862). Chitosan, epigallocatechin gallate (EGCG), and garlic were used as natural bioactives for antibacterial activity. The testing method was carried out according to the disk diffusion method. All of chitosan, EGCG, and garlic showed inhibitory effect against the growth of E. coli and Salmonella typhi. To evaluate the antibacterial activity of natural products during storage, chicken skins were inoculated with $10^6$ of E. coli or Salmonella typhi. The inoculated chicken skins, treated with 0.5, 1, or 2% natural bioactives, were stored during 8 day at $4^{\circ}C$. The numbers of microorganisms were measured at 8 day. Both chitosan and EGCG showed significant decrease in the number of E. coli and Salmonella typhi in dose dependent manner (P < 0.05). These results suggest that natural bioactives such as chitosan, EGCG may be possible to be used as antimicrobial agents for the improvement of food safety.